An Investigation on the Phenotype of Cultured Dendritic Cells from the Peripheral Blood of Patients with Breast Cancer

摘要

Objective To induce and culture the derenditic cells in the peripheral blood of breast cancer patients and research on their phenotypc. Methods Mononuclear cells were isolated by Ficoll Hypaque centrifutation from 32 breast cancer patients' peripheral blood. These cells were plaed in six-well culture plates ( 106 /ml, 2 ml/well) in RPMI 1640 medium supplemented with 10 % heat-in activated fetal bovine serum, 1 00 ng/ml GM-CSF, 20 ng/ml IL-4,and/or 20 ng/ml TNF-a. Two hours later, nonadherent cells were gently removed and fresh medium was added. Cultured cells were ana lyzed by flow cytometry with fluorescence labeled monoclonal antibodies. Pictures of cultured and fluores cence staidned cells were taken by confocal scanning miroscope. Results The diameter of the cells was between 10 and 20 micron. Cells displayed a characteristic CD1a+ ,CD40+ ,CD80+ , CD86+ and CD83+ phenotypes. All of the se molecules were not specific for dendritic cells. CD1a and CD83 molecules could also be expressed on the surface of CD3+ T lymphocyte and CD 19+ B lymphocytes, es pecially on activated lymphocytes. Conclusion The molecules of CD1a and CD83 are not specific phenotypes for dendritic cells. Currently, we still need to apply both cell morphology and costimulatory molecules such as CD40, CD 80, and CD86 to the identificatwn of dendritic cells.